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The study involved the preparation of mixtures of phytic acid ionic liquid and water, which were analyzed for their rheological, frictional, and electrochemical properties. The viscosity and friction characteristics of the mixtures are significantly affected by the water content. Mixtures with low water content (below 30%) result in low friction and negligible wear because of the generation of a close adsorption film and high viscosity. Especially, steel tribo-pairs lubricated with mixtures containing 25% water exhibited friction coefficients lower than 0.06. However, when the water content exceeds 35%, the friction coefficient increases dramatically, and the tribo-surface suffers from severe corrosive wear due to the predominance of the tribo-film in the frictional properties. These results were confirmed through XPS and electrochemical testing.

Monoalkyl phosphates (MAPs) are one kind of important single-chain weak acid/salt type surfactants, but the understanding of their aggregation behavior in water is very limited due to their insolubility at room temperature. In the current work, the effect of guanidinium salts (GuSalts) on the solubility of sodium monododecylphosphate (SDP), a typical MAP, in water was determined at 25.0 °C, and the aggregation behavior of SDP in the GuSalt/water mixtures was investigated. The solubility of SDP is significantly improved by GuSalts including GuCl, GuSO4, GuSO3, GuPO4, and GuCO3 at 25.0 °C, resulting in an isotropic phase. SDP vesicles are spontaneously formed in the isotropic phase, with a critical vesicle concentration of ∼1.0 mM independent of the type of GuSalts. A "bridging dimer" mechanism is proposed to explain the formation of SDP vesicles. The SDP vesicles have a unilamellar structure with a size of ∼80 nm and an alkyl interdigitated degree of ∼25%, and exhibit size-selective permeability. Interestingly, a temperature-induced reversible transition between vesicles and α-gels was observed for the SDP/GuSalt/H2O systems when the SDP content is higher than 20 mM. The α-gels obtained are composed of vesicles and bilayer sheets, showing similar viscoelasticity to conventional gels, although their water content is as high as ∼98 wt%. The microviscosity of SDP vesicle membranes (ca. 35.79-49.34 mPa s at 25.0 °C) and the transition temperature between vesicles and α-gels (ca. 21.0-22.8 °C) are all dependent of the type of GuSalts. This work deepens the understanding of the aggregation behavior of MAPs and also provides valuable information for their practical applications.

Simple single-chain amphiphiles (SCAs) can form vesicular structures in their single-component aqueous solutions, which has attracted great attention, but the understanding of their aggregation behavior is still limited. In this work, the aggregation behavior of 4-dodecylbenzene sulfonic acid (DBSA), a typical simple SCA, in water was investigated. The structure and properties of the aggregates formed were determined. In particular, the effect of wet-dry cycles on the structures of aggregates was examined. The mechanisms of aggregate formation and structural transition were discussed. It was found that the increase of DBSA concentration can drive the occurrence of a micelle-to-vesicle transition, showing a critical micelle concentration and critical vesicle concentration of ∼0.53 and 2.14 mM, respectively. The vesicles formed coexist with micelles in solution, with a unilamellar structure and ∼80 nm size, and exhibit size-selective permeability. In addition, the vesicles show remarkable stability upon long-term storage, exposure to high temperature, and freeze-thaw cycles. The H-bonding interaction between DBSA species and the interdigitated structure of alkyl chains in bilayers play a key role in the formation and stability of DBSA vesicles. Interestingly, it was found that the wet-dry cycle can induce a micelle-to-vesicle transition and an obvious increase in the size of the original vesicles, accompanied by the formation of some multilamellar vesicles. This work provides a better understanding of the aggregation behavior of simple SCAs in their single-component aqueous solutions.

PurposeCollaboration is an important emerging dimension of sustainable supply chain management. How to improve supply chain collaboration (SCC) by means of operational excellence approaches has become an important research topic. The Internet of things (IoT), an important means of operational excellence, has also received increased attention. For better collaboration by the IoT, this study proposes a novel methodology to evaluate the measures of IoT adoption in SCC.Design/methodology/approachBased on the six-domain model and the common classification of collaboration, the measures of the IoT and the criteria of SCC are developed, respectively. A hybrid multi-step methodology that combines neutrosophic set theory, analytic hierarchy process (AHP) and technology for order preference by similarity to an ideal solution (TOPSIS) is proposed to complete the evaluation.FindingsThe results show that improving information transparency, strengthening the integration of management information systems and improving large data processing abilities are the most important measures of the IoT in improving SCC. Measures such as introducing sensing technology and laser scanning technology rank at the bottom and are relatively unimportant.Practical implicationsThe research results provide insights and references for firms to improve SCC by adopting appropriate IoT measures.Originality/valueMost of existing studies indicate the significance of technology in SCC. But this study shows a different conclusion that technologies rank the bottom, while information transparency is more important. And a suitable explanation is given. It further enriches the theoretical studies in SCC field.

Oral lichen planus (OLP) is an inflammatory disease. It is believed that infection and immune dysfunction play a key role in its pathogenesis, but the specific mechanism of action remains unclear. The 16s rRNA high-throughput sequencing technique was used to analyze the microbial flora structure in the saliva of OLP patients and healthy controls. The relative abundance of Derxia, Haemophilus, and Pseudomonas in the saliva of the OLP group was lower than that of the healthy control group, but there was no significant difference in the overall structure of the microbial population. In addition, we measured the protein expression levels of toll-like receptor 4 (TLR4) and nuclear factor-kappab p65 (NF-κB p65) in the tissues of OLP patients, and found that there was a significant increase and positive correlation between them (r = 0.907, P = 0.034). The expression levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in the OLP group were consistent with those of NF-κB p65. Therefore, we believe that changes in the composition ratio of microbialflora break the original balance state of flora, promote the occurrence of immune inflammatory reaction, and then lead to the generation or aggravation of OLP disease. This discovery provides new ideas for further research on OLP initiation and immune regulation mechanism.

The treatment of oral squamous cell carcinoma (OSCC) remains a great clinical challenge, and the malignant proliferation of OSCC cells can lead to the overexpression of CD59. In this study, a novel microsphere (ICT-CMC-CD59sp) composed of icariin (ICT), carboxymethyl chitosan (CMC), and cell differentiation antigen 59-specific ligand peptide (CD59sp) was successfully prepared by using the emulsion cross-linking method. Through the guidance of CD59sp, the microspheres can target OSCC cells and play a therapeutic role (p < 0.01). The MTT test and trypan blue staining showed that the microspheres could promote the apoptosis of oral squamous cell carcinoma and had a significant difference (p < 0.01). In this study, the regulatory effect of the microspheres on OSCC cells was investigated at the cellular level, and its therapeutic effect on OSCC was discussed, which provided a new perspective for the targeted therapy of OSCC.

With the increase of consumption of high-sugar foods, beverages, tobacco, and alcohol, the incidence rate of oral diseases has been increasing year by year. Statistics showed that the prevalence of oral diseases such as dental caries, dental pulpal disease, and periodontal disease has reached as high as 97% in 2015 in China. It is thus urgent to develop functional materials or products for the treatment of oral diseases. Electrospinning has been a widely used technology that is capable of utilizing polymer solution to generate micro/nano fibers under an appropriate high voltage condition. Owing to their excellent structures and biological performances, materials prepared by electrospinning technology have been used for a wide range of oral-related applications, such as tissue restoration, controlled drug release, anti-cancer, etc. In this regard, this article reviews the application and progress of electrospun nanofibers to various oral diseases in recent years. Firstly, engineering strategies of a variety of nanofiber structures together with their resultant functions will be introduced. Then, biological functions of electrospun nanofibers as well as their applications in the treatment of oral diseases are summarized and demonstrated. Finally, the development viewpoint of functional nanofibers is prospected, which is expected to lay the foundation and propose the direction for further clinical application.

Aqueous two-phase systems (ATPSs), consisting of two immiscible water-rich phases, have received great attention. So far, all of ATPSs reported are formed by two water-soluble compounds in aqueous media. Herein, we report an ATPS formed in the single-component aqueous solution of α-ketooctanoic acid (KOCOOH), a weakly acidic surfactant, without any additives. Its formation originates from the coexistence of micelles and vesicles in the system, the former existing in the upper phase and the latter in the lower phase. The phase behavior and microstructures of KOCOOH in aqueous solution were determined. A concentration-driven stepwise aggregation was identified for the KOCOOH solution. With an increase in the KOCOOH concentration, vesicles, oil droplets, micelles, strip bilayers, and planar lamellar phase form successively; macroscopically, the system exhibits a homogeneous transparent single-phase, turbid dispersion, two phases, a bluish single-phase, and a colorless transparent single-phase in turn. The constantly changing ionization state of KOCOOH in aqueous solution plays an important role in the phase and aggregate structure transition. This work deepens the understanding of ATPSs, and the ATPS formed by KOCOOH may have potential applications such as in the separation and purification of biomolecules and the construction of hierarchical protocell models.

Dodecyl hydrogen sulfate (DHS), a simple single-alkyl sulfonic acid, can form vesicles spontaneously in water without any additives close to its apparent pKa. Interestingly, DHS vesicles show typical characteristics of primitive cell membranes in respect of selective permeability, fast exchange, self-reproduction and stability under primordial environments.

Triple-negative breast cancer (TNBC) is a highly aggressive subtype of breast cancer associated with poor prognosis. As an oncogene, DEK involves in regulation of various cellular metabolisms and plays an important role in tumor growth and progression. Increasing evidences suggested that abnormal expression of DEK is closely related to multiple malignant tumors. However, the possible involvement of DEK in epithelial to mesenchymal transition (EMT) and angiogenesis in TNBC remains unclear. In the present study, we revealed that the over-expression of DEK was significantly correlated with clinical stage, differentiation, and lymph node (LN) metastasis of TNBC and indicated poor overall survival of TNBC patients. Moreover, we demonstrated that DEK depletion could significantly reduce cell proliferation, migration, invasion and angiogenesis in vitro. We also found that DEK promoted cancer cell angiogenesis and metastasis by activating the PI3K/AKT/mTOR pathway. Furthermore, we revealed the inhibitory effect of DEK depletion on tumor growth and progression in a xenograft tumor model in mice. These data indicated that DEK promotes TNBC cell proliferation, angiogenesis, and metastasis via PI3K/AKT/mTOR signaling pathway, and therefore, it might be a potential target in TNBC therapy.

Objective To investigate the expression levels of membrane-anchored complement regulatory proteins (mCRPs), CD46, CD55 and CD59 in oral lichen planus (OLP), and evaluate the activation status of complement. Design Thirty-seven cases of OLP patients (20 non-erosive OLP and 17 erosive OLP) and twenty healthy controls were recruited in this study. The proteins and mRNA expression levels of CD46, CD55 and CD59 in OLP tissues were detected by western blotting and RT-qPCR respectively, and the expression levels of complement C3 and sC5b-9 in OLP patients’ saliva were detected by ELISA to evaluate the activation status of complement. In addition, mucosa tissues of another 3 non-erosive OLP patients and another 3 healthy controls were collected, and the epithelial layer of two groups were separated to culture primary keratinocytes in vitro. Immunofluorescence was used to further detect the expression of mCRPs at the cellular level. Results The levels of CD46, CD55 and CD59 in OLP tissues and cells were significantly decreased compared with those of the healthy control group, and the level of complement C3 in the patients’ saliva was significantly decreased, while the level of sC5b-9 was increased. Conclusions These results suggest that the reduced expression of mCRPs keeps the complement system in a continuously active state, which may be the reason of the persistent local immune inflammatory state in OLP. This study aimed to provide new insights for the etiology and therapy of OLP.

The sharing economy has extended to all fields during the last decade, indicating a rapid development. However, many sharing economy firms are facing operational difficulties even bankruptcy, leading to more and more attention have been paid on the sustainability of the sharing economy. Scholars have realized that the collaboration of stakeholders is a potential way to promote the sustainable development of sharing economy. The basis and key rest with understanding the influencing factors of the sharing economy and clarifying the relationships between stakeholders and factors. Only in this way we can put forward specific and practical suggestions based on the research results. Existing studies lack such thinking. Therefore, this paper firstly establishes the value net model of the sharing economy to identify the stakeholders and corresponding influencing factors of the sharing economy. Then a two-mode social network method is used to solve the problems of relationship analysis among different types of elements (stakeholders and factors). The results show that individual suppliers, firm suppliers, and sharing economy platforms are the most influential stakeholders in the sharing economy. The security mechanism, economic benefits, and social motivation are the key factors affecting the sustainability of the sharing economy. Core stakeholders should actively lead and coordinate other stakeholders to establish collaborative relationships and formulate sustainable development strategies based on these key factors.

The phase behavior of sodium monododecylphosphate (SDP), a typical primitive single-chain amphiphile, in n-butanol (NBT)/H2O mixed solvent was investigated. An isotropic phase was observed in the SDP/NBT/H2O ternary system. Three aggregation structures, namely, vesicles, dendritic aggregates, and micelles, were identified in the isotropic phase. The SDP vesicles were characterized by transmission electron microscopy, dynamic light scattering, atomic force microscopy, and small-angle X-ray scattering. Their stability and permeability were determined. In addition, two kinds of enzyme-loaded vesicles were established to support a cascade reaction, to explore the possibility for chemical signaling. The results showed that the critical vesicle concentration of SDP in the solvent is ∼0.30 mM. The size and shell thickness of the vesicles are ∼80 and 3.81 nm, respectively. The vesicle formation probably arises from the synergistic effect of the solubilization of NBT for SDP and the H-bonding between SDP species. The SDP vesicles exhibit long-term and freeze-thawing stabilities and a size-selective permeability. In particular, the SDP vesicles can host a cascade reaction to mimic a chemical-signaling communication scenario, which may be a suitable alternative to protocell models.

Background T-acute lymphoblastic leukemia (T-ALL) was a hematological malignancy characterized by the accumulation of immature T cells in bone marrow and peripheral blood. In this study, we tried to explore the physiological role of CD59 in T-ALL. Methods In this study, we collected the bone marrow samples from 17 T-ALL patients and 38 healthy participants to find differences in CD59 expression patterns. Then, CD59 was over-expressed in T-ALL cell line Jurkat, and its biological functions were detected. In addition, in order to understand the active site of CD59, the Trp40 was mutated. Further, we constructed a mouse model by transplanting Jurkat cells into the nude mice to verify the function of CD59 in vitro. At last, mechanism studies were performed by western blot. Results We found that the proportion of T lymphocytes expressing CD59 in bone marrow of T-ALL patients was significantly higher than that of healthy individuals. Then, we found that the overexpression of CD59 in Jurkat cells was beneficial to the cell survival by inhibiting apoptosis and promoting IL-2 secretion. In this process, Trp40 of CD59 was a key functional site. Further, the high expression of CD59 inhibited apoptosis of bone marrow and peripheral blood cells, and promoted IL-2 secretion in mouse model. At last, mechanism studies showed that the activation of AKT, STAT5 and Notch1 signaling pathways in Jurkat cells, may be involved in the regulation of apoptosis by CD59; and mutation in the Trp40 affect the interaction of CD59 with these signaling pathways. Conclusions In conclusion, CD59 inhibited apoptosis of T-ALL by regulating AKT/Notch1 signaling pathway, providing a new perspective for the treatment of T-ALL. Electronic supplementary material The online version of this article (10.1186/s12935-018-0714-9) contains supplementary material, which is available to authorized users.

Photodynamic therapy (PDT), which is a procedure that uses photosensitizing drug to apply therapy selectively to target sites, has been proven to be a safe treatment for cancers and conditions that may develop into cancers. Nano-sized TiO2 has been regarded as potential photosensitizer for UV light driven PDT. In this study, four types of TiO2 nanofibers were prepared from proton tri-titanate (H2T3O7) nanofiber. The as-obtained nanofibers were demonstrated as efficient photosensitizers for PDT killing of HeLa cells. MTT assay and flow cytometry (FCM) were carried out to evaluate the biocompatibility, percentage of apoptotic cells, and cell viability. The non-cytotoxicity of the as-prepared TiO2 nanofibers in the absence of UV irradiation has also been demonstrated. Under UV light irradiation, the TiO2 nanofibers, particularly the mixed phase nanofibers, displayed much higher cell-killing efficiency than Pirarubicin (THP), which is a common drug to induce the apoptosis of HeLa cells. We ascribe the high cellkilling efficiency of the mixed phase nanofibers to the bandgap edge match and stable interface between TiO2(B) and anatase phases in a single nanofiber, which can inhibit the recombination of the photogenerated electrons and holes. This promotes the charge separation and transfer processes and can produce more reactive oxygen species (ROS) that are responsible for the killing of HeLa cells.

CD59 may inhibit the cytolytic activity of complement by binding to C8/C9 and protect host cell membranes against homologous membrane attack complex (MAC). However, CD59 is widely overexpressed on tumor cells, which has been implicated in tumorigenesis. The active site of CD59 relative to MAC is still confused. As reported the MAC binding site is located in the vicinity of a hydrophobic groove on the membrane distal face of the protein centered around residue W40. Here two site-directed mutagenesis were performed by overlapping extension PCR to delete residue W40 site (Mutant 1, M1) or to change C39W40K41 to W39W40W41 (Mutant 2, M2). Then we constructed mutant CD59 eukaryotic expression system and investigated their biological function on CHO cells compared with wild-type CD59. Stable populations of CHO cells expressing recombinant proteins were screened by immunotechnique. After 30 passages culturing, proteins could be tested. Dye release assays suggest that M1CD59 loses the activity against complement, while M2CD59 increases the anti-complement activity slightly. Results indicate that W40 of human CD59 is important to its activity, and prohibition of this site may be a potential way to increase complement activity and to treat tumors.

To establish xenograft mouse models of Jurkat T-leukemia cells over-expressing C-terminal Src kinase binding protein (CBP).The 5-week-old female BALB/c-nu mice were randomly divided into blank control group, normal Jurkat cell control group, empty virus-transfected Jurkat cell control group and CBP over-expression model group, 5 mice in each group. The mice were subcutaneously injected 1×10(7)/0.1 mL Jurkat cells in axillary area. The tumor tissues of mouse models were weighed and then subjected to HE staining to observe the pathological changes of tumor tissues. The proliferation of Jurkat cells in the peripheral blood of mice was detected by flow cytometry, and the interleukin 2 (IL-2) levels in mouse sera were determined with ELISA.The volume of tumor tissues in the CBP over-expression model group was smaller than that in the control groups, so was the mass of tumor tissues. HE staining showed the proliferation of Jurkat cells in tumor tissues of the model group. The proliferation rate of Jurkat cells in the peripheral blood and IL-2 levels in the sera of the CBP over-expression model group were lower than those in the normal Jurkat cell control group and the empty virus-transfected Jurkat cell control group.The mouse models of Jurkat T-leukemia cells over-expressing CBP have been established successfully. Up-regulated CBP has inhibitory effects on the proliferation of Jurkat cells and IL-2 secretion.

To construct the lentivirus carrying the mutated palmitoylation site of the linker for activation of T cells (LAT) and infect Jurkat cells with it to establish stable cell line, and to investigate the effect of LAT palmitoylation mutation on T cell signaling induced by CD59.Negative control (neg-EGFP) and LAT-M-EGFP fusion protein gene vectors were respectively constructed and then packaged using lentivirus. Subsequently, Jurkat cells were infected with them to establish stable cell lines. Confocal laser scanning microscopy was used to observe the infection efficiency and the distribution of fusion proteins in Jurkat cells. CCK-8 assay was used to detect the change of cell proliferation activity after CD59 mAb supplementation. Flow cytometry was used to determine the apoptosis rate. Western blotting was used to examine the levels of phospholipase C-γ1 (PLC-γ1) and lymphocyte-specific protein tyrosine kinase (LCK).Confocal laser scanning microscopy revealed that LAT molecules of LAT-M group scattered on cell membrane, and there was no obvious clustered region after cross linkage with CD59 mAb. Compared with the negative control group, the cell proliferation activity of LAT-M group significantly decreased, and the quantity of middle-late apoptotic cells significantly increased; Western blotting showed that the expression levels of PLC-γ1 and LCK in LAT-M group was roughly the same with those in negative control group, and after CD59 mAb stimulation, there was no obvious change in LAT-M group, while the levels in negative control group were reduced.LAT-M-EGFP fusion protein could not locate on lipid rafts of Jurkat cells infected with LAT palmitoylation mutation. In addition, the growth of the cells carrying the LAT-M-EGFP was inhibited. The palmitoylation mutation of LAT attenuated the signal transduction induced by glycosylphosphatidylinositol-anchored CD59 in T cells.

To explore the effects of up-regulated expression of C-Src kinase-binding protein (CBP) on biological functions and cell ultrastructure in Jurkat cells.We constructed the CBP-EGFP viral vector and established a stably transfected Jurkat cell line with it. Cell growing status was observed under a light microscope. The transfection efficiency and the location of CBP were examined with confocal microscopy. The complexity of cell organelles was observed by electron microscopy. Apoptotic cells were detected by flow cytometry. The expressions of C-Src kinase (CSK) and Vav oncoprotein in CBP signaling pathway were measured by real-time quantitative PCR (qRT-PCR).We found more pyknotic cells and less homogeneity in the transfected Jurkat cells under light microscope. Confocal microscopy revealed that cell transfection efficiency was up to 100% and CBP was located on the cell membrane. Moreover, there were more complex organelles in CBP-EGFP transfected Jurkat cells with a lot of mitochondrions and the lysosome-like clumps accumulation. Detection of apoptosis showed that CBP-EGFP transfected Jurkat cells had more necrotic cells compared with the negative group. Finally, qRT-PCR indicated the expression of CSK increased and Vav decreased in the CBP-EGFP transfected group.Up-regulated expression of CBP in Jurkat cells could reduce cell homogeneity and promote cell apoptosis.

Item does not contain fulltext Von Recklinghausen's disease is a relatively common familial genetic disorder characterized by inactivating mutations of the Neurofibromatosis-1 (NF1) gene that predisposes these patients to malignancies, including an increased risk for juvenile myelomonocytic leukemia. However, NF1 mutations are not common in acute myeloid leukemia (AML). Given that the RUNX1 transcription factor is the most common target for chromosomal translocations in acute leukemia, we asked if NF1 might be regulated by RUNX1. In reporter assays, RUNX1 activated the NF1 promoter and cooperated with C/EBPalpha and ETS2 to activate the NF1 promoter over 80-fold. Moreover, the t(8;21) fusion protein RUNX1-MTG8 (R/M), which represses RUNX1-regulated genes, actively repressed the NF1 promoter. R/M associated with the NF1 promoter in vivo and repressed endogenous NF1 gene expression. In addition, similar to loss of NF1, R/M expression enhanced the sensitivity of primary myeloid progenitor cells to granulocyte-macrophage colony-stimulating factor. Our results indicate that the NF1 tumor suppressor gene is a direct transcriptional target of RUNX1 and the t(8;21) fusion protein, suggesting that suppression of NF1 expression contributes to the molecular pathogenesis of AML.

Correction for ‘Model of protocell compartments – dodecyl hydrogen sulfate vesicles’ by Bin Liu et al., Phys. Chem. Chem. Phys., 2018, DOI: 10.1039/c7cp06379b.

To intervene the expression of CD59 on cervical cancer HeLa cells by RNA interference and phage display random peptide library, respectively, and detect the effect of down-regulated CD59 expression on proliferation and apoptosis of the cervical cancer cells.HeLa cells were divided into normal control group, CD59 peptide seal group, pSUPER transfected group and pSUPER-siCD59 transfected group. The CD59 peptide seal group was treated with 10 μg/mL CD59 peptide seal for 8 hours. The pSUPER and pSUPER-siCD59 transfected groups were transfected with empty plasmid pSUPER and recombinant plasmid pSUPER-siCD59, respectively. MTT assay was used for the detection of cell proliferation. TUNEL, annexin V-PE/7-AAD staining combined with flow cytometry were adopted for the detection of cell apoptosis.Compared with the control groups, both CD59 peptide seal group and pSUPER-siCD59 transfected group presented a diminished cell proliferation activity and a strengthened apoptosis. What's more, the effect of peptide seal on CD59 expression inhibition was better than that of pSUPER-siCD59 plasmid.Down-regulation of CD59 expression could inhibit HeLa cells' proliferation and promote apoptosis, and the inhibitory effect of peptide seal was better than that of CD59 interference plasmid.

CD59 is a crucial complement regulatory protein that inhibits the terminal step of the complement activation cascade by interfering with the binding of C9 to C5b-8, thus preventing the formation of the membrane attack complex (MAC). We recently reported that the mouse genome contains two Cd59 genes, while the human and rat genomes each contain only one Cd59 gene (Qian et al. 2000). Here, we describe the genomic structure, comparative activity, and tissue distribution of these two mouse genes, designated Cd59a and Cd59b. The mouse Cd59 genes encompass a total of 45.6 kb with each gene having four exons. Cd59a spans 19 kb, and Cd59b spans 15 kb, with approximately 11.6 kb of genomic DNA separating the two genes. The overall sequence similarity between Cd59a and Cd59b is approximately 60%. The sequence similarity between exon 2, exon 3, and exon 4 and the respective flanking regions between the two genes is over 85%, but exon 1 and its flanking regions are totally different. Comparative studies of the activity of both genes as inhibitors of MAC formation revealed that Cd59b has a specific activity that is six times higher than that of Cd59a. Using polyclonal antibodies specific to either Cd59a or Cd59b, we showed that Cd59a and Cd59b are both widely expressed in the kidneys, brain, lungs, spleen, and testis, as well as in the blood vessels of most mouse tissues. Interestingly, testicular Cd59a appeared to be expressed exclusively in spermatids, whereas Cd59b was expressed in more mature sperm cells. These results suggest that even though Cd59a and Cd59b are expressed in multiple tissues, they may play some different roles, particularly in reproduction.

We have previously shown that left ventricular (LV) pacing-induced heart failure is associated with preserved wall thickening in the interventricular septum (IVS) compared with the posterolateral wall (PLW). The current study focuses on the relationship between regional myocardial function and altered β-adrenergic receptor (β-AR) signaling. We studied 15 pigs: 6 controls and 9 paced from the left ventricle (225 beats/min, 26 ± 3 days). Heart failure was documented by decreased LV fractional shortening ( P < 0.0001) and increased left atrial pressure ( P < 0.0001). In heart failure, despite marked differences in basal regional function (percent wall thickening: IVS, 33 ± 10% vs. PLW, 13 ± 7%; P = 0.0003), there were no differences between the two regions in β-AR responsiveness, measured by regional wall thickening in response to dobutamine infusion and any measurement of adrenergic signaling. Adenylyl cyclase activity, β-AR number, and β-AR/Gscoupling were markedly reduced in failing LV without regional differences. In animals with heart failure, LV G protein receptor kinase (GRK) isoform 2 content was unchanged and GRK5, the other major GRK isoform, was increased more than threefold (IVS, 0.51 ± 0.20 vs. 0.12 ± 0.12 arbitrary densitometric units, P = 0.01; PLW, 0.47 ± 0.15 vs. 0.13 ± 0.09 arbitrary densitometric units, P = 0.03), but again, there were no regional differences. These data indicate that systemic rather than regional factors govern LV adrenergic signaling and that regional adrenergic signaling abnormalities poorly predict wall thickening in the same regions.

The solid state photochemistry of uranyl carboxylate complexes is presented with the purpose of developing methods for optical lithography of uranium oxide films. These complexes of the general formula, UO2(OOCR)2 (R = i-C3H7, C5H11, CH2C6H5, CH2OC2H5, C2H4OC2H5), were all photosensitive as thin amorphous films. The primary photochemical reaction for each of these complexes was the extrusion of a CO2 from the ligand and the production of radicals which initiated a chain reaction. The nature of this chain reaction was dependent upon the identity of the organic substituents, R. In some cases the chain reaction required a photochemical step while others were entirely thermal in nature. Of importance are the potentially high quantum yields which can be associated with thermal chain reactions. Some of the systems presented here exhibit quantum yields in excess of 1. This process was shown to be compatible with optical lithography by the patterning of the uranium oxide product on silicon surfaces.

Cellular content of cAMP generated by activation of adenylylcyclase (AC; EC 4.6.1.1 ) is a key determinant of functional responsiveness in the heart and other tissues. We have tested two hypotheses regarding the relationship between AC content and β-adrenergic receptor (βAR)-mediated signal transduction in cardiac myocytes. First, that AC content limits adrenergic signal transduction, and, second, that increased AC, independent of (βAR) number and G-protein content, yields a proportional increase in βAR-mediated transmembrane signaling. We used recombinant adenovirus to increase AC isoform VI (AC VI ) expression in neonatal cardiac myocytes. Cells that overexpressed AC VI responded to agonist stimulation with marked increases in cAMP production in proportion to protein expressed. In parallel experiments performed on cells transfected with lacZ (control) or AC VI , [ 3 H]forskolin binding, used to assess AC protein expression, was amplified 6-fold, while βAR-stimulated cAMP production from these cells was increased 7-fold. No changes in βAR number, or in the heterotrimeric GTP-binding proteins, Gαs or Gαi 2 , were observed. Previous studies indicate that increased cardiac expression of βAR or Gαs does not yield proportional increases in transmembrane adrenergic signaling. In contrast, the current data demonstrate that increased AC VI expression provides a proportional increase in β-adrenergic signal transduction. Our results show that the amount of AC sets a limit on transmembrane β-adrenergic signaling. We speculate that similar functional responses are possible in other cell types in which AC plays an important physiological role.

To transfect Jurkat cells with linker for activation of T cells (LAT)-EGFP or LAT-EGFP-M (two cysteines of which near the juxtamembrane region were substituted for alanine) by electroporation, and observe the change in T cell signal transduction after the cross linkage with anti-CD59 mAb.By means of electroporation, Jurkat cells were transfected with LAT-EGFP or LAT-EGFP-M or PEGFP-N3 (control plasmid), and then subjected to cross linkage with anti-CD59 mAb. The locations of the fusion protein LAT-EGFP and LAT-EGFP-M were observed with confocal microscopy. The proliferation activity of the Jurkat cells was determined by MTT assay, and the content of interleukin 2 (IL-2) in cell supernatants was detected by ELISA. Western blotting was used to test the phosphorylation level of LAT in each group.After the cross linkage of anti-CD59 mAb, LAT-EGFP-M could not locate on lipid rafts just as LAT-EGFP did. The cell proliferation and IL-2 secretion of Jurkat cells transfected with LAT-EGFP-M decreased as compared with the ones transfected with LAT-EGFP, EGFP-N3 or the untransfected ones. The phosphorylation level of LAT-EGFP-M was much lower than that of LAT-EGFP.The mutation of palmitoylation site of LAT-EGFP attenuated the signal transduction of CD59 in T cells.

To investigate the distributions of GPI-anchored protein CD59 and C-terminal Src kinase-binding protein (Cbp) in cell membrane of T lymphocytes and the roles in cell activation and proliferation.The locations of CD59 and Cbp were observed under a fluorescence microscope with the immunofluorescence cytochemistry. The Jurkat cells were transfected with the recombinant plasmid pSUPER-siCD59 using the electric transfection method. Using RT-PCR and Western blotting, we detected the expression of CD59 and the phosphorylation level of protein-tyrosine kinase oncogene family (fyn) in different groups. Moreover, the cell proliferation activity was measured by MTT assay.The immunofluorescence cytochemistry showed that CD59 and Cbp were mainly distributed in cell membrane. Compared with other groups, the quantitative RT-PCR and Western blotting showed that the expression of CD59 was reduced, and along with it the phosphorylation level of fyn decreased in pSUPER-siCD59 group, and MTT assay revealed that its cell proliferation was inhibited (P0.05).CD59 was a type of membrane-bound protein, CD59 and Cbp played synergistic roles in cell activation and proliferation.

The photolysis of the complexes UO 2 (OH 2 )(RCOCHCOR) 2 (R  Me or t -C 4 H 9 ) on silicon surfaces leads to the production of uranium oxide. The quantum yields for this process are quite different for the two complexes. The methyl and the tert -butyl-substituted complexes decompose with quantum yields of 0.007 (±40%) and 0.015 (±30%) respectively. In spite of the similarity in chemical formula a detailed mechanistic investigation indicates that two distinct reaction pathways exist for these systems. In the case of the methyl derivative the reactivity is initiated by an excited state hydrogen abstraction through a six-member transition state. The tert -butyl-substituted complex cannot undergo hydrogen abstraction and so the excited state decays by a ligand fragmentation reaction.

The photochemistry of coordination complexes in the solid state has been investigated with a view towards developing a new method for the photolithographic deposition of materials. Ultraviolet exposure of thin films of coordination complexes through an optical lithography mask results in the patterning of materials in the exposed regions. The development of this method requires photochemically active complexes which efficiently eject ligands upon photolysis to produce the desired material in a solid state thin film. A generic reaction for the production of metal films is M L n ( thin film ) → h v M ( thin film ) + n L ( g ) In this paper the photochemistry of thin films of inorganic complexes of Co, Cu, Ni, Pd, Pt and U is presented. These examples will be used to outline the approaches used to design suitable precursor molecules for film deposition. Mechanistic studies of the surface photochemical reactions along with evidence for submicron resolution lithography of materials derived from these complexes will be highlighted. The use of electron beams to induce similar chemistry will also be discussed.

Apolipoprotein E (ApoE), which has been shown to influence serum lipid parameters, can bind to multiple types of lipids and plays an important role in the metabolism and homeostasis of lipids and lipoproteins. A previous study showed that ApoE concentration significantly affects serum lipid levels independently of ApoE polymorphism. The serum lipid levels were also closely correlated with dietary habits, and Shandong cuisine is famous for its high salt and oil contents, which widely differ among the different areas in China. Therefore, studying the effect of ApoE polymorphism on ApoE concentration and serum lipid levels in Shandong province is very important. A total of 815 subjects including 285 men and 530 women were randomly selected and studied from Jinan, Shandong province. In order to evaluate the association of ApoE polymorphism and serum level on lipid profiles, the ApoE genotypes, as well as levels of fasting serum ApoE and other lipid parameters, were detected in all subjects. The frequency of the ApoE E3 allele was highest (83.1%), while those of E2 and E4 were 9.4% and 7.5%, respectively, which are similar to those in other Asian populations. ApoE2 allele carriers showed significantly increased ApoE levels but lower levels of serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and Apolipoprotein B (ApoB). We found that ApoE level is influenced by ApoE polymorphism in a gene-dependent manner. The ApoE polymorphism showed different influences on serum lipid parameters with increasing age and body mass index (BMI) in our Shandong Han population.

The aim of this study was to investigate the pro-apoptotic mechanism of C-phycocyanin (C-PC)-mediated photodynamic therapy (PDT) in a murine tumor model and cultured MCF-7 cells. The mice were divided into four groups: control, He-Ne laser radiation, C-PC treatment, and C-PC treatment + He-Ne laser radiation. The effects of C-PC and/or laser on immune organs, immunocyte proliferation, tumor genesis, and apoptosis-related proteins expressions were investigated by immunohistochemistry, in situ hybridization, MTT, electron microscope, western blot, and immunofluorescence assay. The results showed that He-Ne laser treatment alone showed marginal effects. In C-PC-treated mice, the weight of immune organs, proliferation of immunocytes, and expression of pro-apoptotic Fas protein were increased, whereas the tumor weight and the expressions of anti-apoptotic proteins (NF-kappaB and P53) and CD44 mRNA were comparatively decreased. In vitro, C-PC was able to inhibit MCF-7 cell proliferation and cause ultrastructural changes including microvilli loss, formation of membrane blebs, and chromatin condensation. Moreover, C-PC treatment could activate caspase-9 expression, induce cytochrome c release, and downregulate Bcl-2 expression. When combined with He-Ne laser irradiation, the effects of C-PC treatment were further enhanced. Facilitating the apoptosis signals transduction and finally leading to the apoptosis of MCF-7 cells may be the mechanism of the anti-tumor activities of C-PC-mediated PDT.

Vascular proliferation is one of the major causes for morbidity and mortality in diabetes. However, the cellular and molecular mechanisms that link hyperglycemia to this complication remain unclear. In present study, we demonstrated by site-directed mutagenesis that mutated CD59 was more susceptible to glycation-inactivation for hyperglycemia. Mutated and wild-type CD59s were stably expressed in Chinese hamster ovary cells using the pALTER-MAX mammalian expression vector. Western blot, FACS and immunological fluorescence were conducted to confirm that CD59s were tethered to the plasma membrane. Compared to wild-type CD59, human CD59 mutants led to a significant increase in dye release assay. These results indicate that there may be some mutations of CD59 in diabetes population and the mutated CD59, which is more likely to be of glycation than the wild-type, may help to explain the distinct propensity of diabetes subjects to develop vascular proliferation complications.

This study is aimed at investigating the significance of gene promoter methylation status and protein expression of p16 INK4A and O6-methylguanine-DNA methyltransferase (MGMT) in the various uterine cervical lesions. Methylation status by using methylation-specific polymerase chain reaction (MS-PCR) and protein expression by using immunohistochemistry for p16 INK4A and MGMT genes were performed in cervical squamous intraepithelial neoplasms (CIN), invasive squamous cell carcinomas (SCC), adenocarcinomas and non-neoplastic cervices. None of 20 non-neoplastic cervices showed p16 INK4A and MGMT gene hypermethylation, whereas at least one of these genes was hypermethylated with 50.0% (5/10) of CIN I, 65.0% (13/20) of CIN II–III, 70.2% (33/47) of SCC and 85.0% (17/20) of adenocarcinoma. p16 INK4A protein was totally negative in non-neoplastic cervices, but positive with 90.0% of CIN I, 100% of CIN II–III and adenocarcinoma, and 78.7% of SCC. MGMT protein was expressed in 10% of non-neoplastic cervices, but significantly increased in SCC (42.5%) and adenocarcinoma (70.0%). The protein expression of p16 INK4A and MGMT was not related to their gene promoter methylation status. The hypermethylation of p16 INK4A and MGMT genes in the uterine cervix may indicate the presence of malignant cells, and p16 INK4A immunostaining is useful in grading CIN and diagnosing invasive SCC and adenocarcinoma.